Magnetic resonance spectroscopy (MRS) is a preeminent diagnostic method in medical science for depicting chemical composition, disease, and function. The advancement of more powerful and precise MRS instrumentation is critical to broaden the applicability of the technology to biomedical research and disease diagnosis, especially as it relates to small-scale studies including in-cell spectroscopy. Here, we test the hypothesis that novel radiofrequency microcoils fabricated on the cantilever portion of standard atomic force microscopy (AFM) probes can enable a new hybrid AFM/MRS technology. We use nanofabrication processes developed in our laboratory to create planar RF coils formed on a deflectable AFM sensing probe, therefore allowing for highly-focused spectroscopy and nanometer-discretized cell localization and biophysical analysis. We will pursue two Aims. In Aim 1, we will optimize the detection resolution and performance of AFM-coupled micro radiofrequency (?RF) probes. Probes will be evaluated in hydrogel systems with known relativity. In Aim 2, we will determine the extent that ?RF probes enable noninvasive spectroscopy of single cells. We will conduct MRS studies on cell systems through the implementation of a piezoelectric-based feedback mechanism for the deflection measurement of the AFM tip, which takes place when the tip is brought into contact with samples. Successful completion of the proposed experiments is expected to provide researchers with a new tool that will compete with existing diagnostic tools in terms of localization precision of morphological and chemical data and enable analysis of subsurface features and biophysical investigation in the cellular level. PUBLIC HEALTH RELEVANCE: We aim to complement the biochemical analysis capabilities of high-resolution magnetic resonance spectroscopy (MRS) with the spatial localization features and biophysical analysis provided by atomic force microscopy (AFM), through the development of a new instrumentation tool for real-time spectroscopy, biophysical, and structural analysis of single cells. MRS is a preeminent analytical method for the acquisition of chemical and structural information within molecules. With the detection of biomarkers through the employment of amplification techniques, combined with AFM-enhanced localization, MRS can be used as a diagnostic tool, especially with the development of advanced in-cell MRS reconstruction techniques.